Multiplex Detection of Aspergillus fumigatus Mycoviruses.

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).Mycoviruses are viruses that naturally infect and replicate in fungi. They are widespread in all major fungal groups including plant and animal pathogenic fungi. Several dsRNA mycoviruses have been reported in Aspergillus fumigatus. Multiplex polymerase chain reaction (PCR) amplification is a version of PCR that enables amplification of different targets simultaneously. This technique has been widely used for detection and differentiation of viruses especially plant viruses such as those which infect tobacco, potato and garlic. For rapid detection, multiplex RT-PCR was developed to screen new isolates for the presence of A. fumigatus mycoviruses. Aspergillus fumigatus chrysovirus (AfuCV), Aspergillus fumigatus partitivirus (AfuPV-1), and Aspergillus fumigatus tetramycovirus-1 (AfuTmV-1) dsRNAs were amplified in separate reactions using a mixture of multiplex primer pairs. It was demonstrated that in the presence of a single infection, primer pair mixtures only amplify the corresponding single virus infection. Mixed infections using dual or triple combinations of dsRNA viruses were also amplified simultaneously using multiplex RT-PCR. Up until now, methods for the rapid detection of Aspergillus mycoviruses have been restricted to small scale dsRNA extraction approaches which are laborious and for large numbers of samples not as sensitive as RT-PCR. The multiplex RT-PCR assay developed here will be useful for studies on determining the incidence of A. fumigatus mycoviruses. This is the first report on multiplex detection of A. fumigatus mycovirusesPeer reviewe

Are BPA-free plastics safe for aquatic life? - Fluorene-9-bisphenol induced thyroid-disrupting effects and histopathological alterations in adult zebrafish (Danio rerio)

Fluorene-9-bisphenol (BPFL) is used as an alternative compound for bisphenol A, an endocrine disruptor compound which is present in various materials including plastic bottles and packaging. Although it is used extensively in products that are labelled BPA-free, its effect on wildlife and humans have not been fully studied. Therefore, this study aimed to investigate the effects of BPFL in adult zebrafish. In the preliminary experiments of the study, the median lethal concentration value (LC50) of BPFL was 0.25 mg/L (95 % confidence interval 0.15–0.41) for 96 h. Following exposure to three different sublethal concentrations of BPFL after 96 h and 15 days, T4 hormone levels, expression levels of genes involved in thyroid metabolism and histopathological alterations were assessed. T4 hormone levels were found to be significantly higher in females at the lowest BPFL concentration following 96 h exposure (P < 0.05). Expression levels of trh, tshba and trhrb genes were upregulated following 96 h exposure at 0.025 mg/L concentration and crh was upregulated following 15 days exposure at 0.025 mg/L concentration in female zebrafish (P < 0.05). The most prominent histopathological findings in zebrafish exposed to 0.025 and 0.125 mg/L of BPFL were observed in the gill, liver, kidney and testis tissues. The gill tissues showed some hyperemia, lamellar fusion, hyperplasia, epithelial lifting, and telangiectasis, while passive hyperemia, hydropic degeneration, and necrosis were observed in the liver tissues. The BPFL is highly toxic to zebrafish even in sublethal concentrations according to the molecular and histopathological responses. © 2022 Elsevier Inc

Sublingual buprenorphine/naloxone treatment is not affected by OPRM1 A118G and BDNF Va66Met polymorphisms, but alters the plasma beta-endorphin and BDNF levels in individuals with opioid use disorder

The study aimed to examine the genetic contribution to buprenorphine (BUP) treatment in individuals with opioid use disorder (OUD), with a specific focus on BDNF and OPRM1 genes. A total of 113 controls and 111 OUD patients receiving sublingual BUP/naloxone were enrolled. OPRM1 A118G and BDNF Val66Met polymorphisms were investigated by PCR-FRLP. Plasma BDNF and beta-endorphin levels were assessed by ELISA kits in both groups. Blood BUP levels were measured by LC-MS/MS and normalized with daily BUP dose (BUP/D). OPRM1 A118G and BDNF Val66Met polymorphisms didn't have an effect on plasma beta-endorphin and BDNF levels in OUD patients, respectively. Interestingly, OUD patients had significantly higher plasma BDNF and lower beta-endorphin levels compared to the controls (p < 0.001). A negative and significant correlation between plasma BUP/D and BDNF levels was found. Age onset of first use was associated with OPRM1 A118G polymorphism. The findings indicated that sublingual BUP/naloxone may increase plasma BDNF levels, but may decrease beta-endorphin levels in individuals with OUD. Plasma BDNF level seemed to be decreased in a BUP/D concentration-dependent manner. © 2022 Elsevier B.V

Tatlısu ekosisteminde endişe yaratan kirleticiler: Türkiye’den bir vaka çalışması

The current study evaluated certain emerging contaminants in the Susurluk sub-basin, an area under significant anthropogenic pollution pressure. Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), organochlorine insecticides, and microplastics were investigated both from surface and sediment samples collected in dry and wet seasons. γ-HCH and β-HCH were detected in the dry season from Kocaçay River reaching the Marmara Sea. Dieldrin concentrations were also very high in river sediments during the dry season. Nilüfer Stream is a significant waterway close to industrial and urban areas and under impact of pollution due to high concentrations of PCBs, DDT, and its’ metabolites. Sediment samples contained higher levels of contaminants: DDT and metabolites were found in sediments from almost all stations. According to the Hazard quotient coefficient, all detected pollutants were found to be >1, indicating a high risk in the river system. Fiber was the dominant microplastic. The water quality of Nilüfer Stream was poor/bad in quality both in dry and wet seasons, while Kocaçay River was moderate and poor/bad quality in dry and wet seasons, respectively. The findings of bacterial growth augmented and worsened water quality in the river basin with coliforms dominating, as assessed at the genus/species level and were very abundant.Bu çalışma, önemli antropojenik kirlilik baskısı altındaki bir alan olan Susurluk alt havzasında ortaya çıkan bazı endişe yaratan kirleticiler değerlendirmiştir. Kurak ve yağışlı mevsimlerde toplanan yüzey ve sediman örneklerinde poliklorlu bifeniller (PCBler), polibromlu bifeniller difenil eterler (PBDEler), organoklorlu insektisitler ve mikroplastikler araştırılmıştır. γ-HCH and β-HCH, Kocaçay Nehri’nin Marmara Denizi’ne dökülen bölgesinde kurak mevsimde tespit edilmiştir. Dieldrin konsantrasyonları da kurak mevsim boyunca nehir sedimanlarında çok yüksektir. Nilüfer çayı endüstriyel ve kentsel alanlara yakın olan önemli bir su yoludur ve yüksek konsantrasyondaki PCBler, DDT ve metabolitleri nedeniyle kirlilik etkisi altındadır. Sediman örnekleri daha yüksek seviyede kirletici içermektedir: DDT ve metabolitleri neredeyse tüm istasyonlardan alınan sediman örneklerinde bulunmuştur. Tehlike oranı katsayısına (Hazard Quotient) göre tesbit edilen tüm kirleticiler >1 olarak bulunmuş ve bu da nehir sisteminde yüksek risk olduğunu göstermiştir. Mikroplastikler içinde fiber en baskın olanıdır. Nilüfer Çayı’nın su kalitesi hem kurak hem de yağışlı sezonda kötü iken, Kocaçay Nehri’nin su kalitesi kurak mevsimde orta ve yağışlı mevsimde kötüdür. Cins/tür düzeyinde değerlendirildiğinde koliformların hakim olduğu ve çok miktarda olan bakteriyel büyüme bulguları da nehir havzasındaki su kalitesini kötüleştirmiştir

Optimization of PCR-RFLP method for genotyping of GHSR rs2232165 Gene Polymorphism and Determination of allele frequency in Turkish men

Çağımızın ciddi sorunlardan biri olan alkol bağımlılığı, genetik, çevresel, kültürel, gelişimsel ve nörobiyolojik faktörlerin etkisiyle ortaya çıkmaktadır. Son yıllarda yeme davranışı nörobiyolojisi ile bağımlılık yapan maddelere aşerme davranışının nörobiyolojisinin benzer özelliklere sahip olduğunun keşfedilmesi araştırmacıları, alkol bağımlılığı ile grelin gibi besin alımında önemli olan hormonlar arasındaki ilişkiyi araştırmaya sevk etmiştir. Alkol kullanım bozukluğunda ve alkole aşermede grelin sisteminin önemi mevcut çalışmalar tarafından ortaya konulmasına karşın bu ilişkinin biyolojik mekanizmasının aydınlatılması için daha çok çalışmaya ihtiyaç vardır. Bu nedenle, bu çalışmada, alkol kullanım sorunu olan Türk erkeklerinde, grelin reseptörünü kodlayan GHSR genindeki rs2232165 polimorfizminin alkol kullanım sorunu ile ilişkisi araştırılmıştır. Çalışmaya alkol kullanım bozukluğu tanısı konmuş 72 erkek birey ile herhangi bir madde bağımlılığı olmayan 82 sağlıklı erkek dahil edilmiştir. GHSR rs2232165 gen polimorfizmi, ilk defa bu çalışmada optimize edilen PCR-RFLP yöntemi ile genotiplendirilmiştir. GHSR rs2232165 polimorfizmi için alel frekansları alkol kullanım sorunu olan grupta (n:144); C aleli için 0,99 (n:142), T aleli için 0,01 (n:2) olarak hesaplanmıştır. Karşılaştırma grubunda ise (n:164), C aleli frekansı 0,98 (n:161), T aleli frekansı 0,02 (n:3) olarak belirlenmiştir. İki grup arasında, T alel frekansı açısından istatistiksel olarak anlamlı bir fark bulunmamıştır (p>0,05). Gen polimorfizmleri, çevresel faktörlerden etkilenmediği için Türk erkeklerindeki minör alel frekansını (MAF) belirlemek için iki gruptaki bireyler birleştirilmiş (n=154) ve Türk erkeklerinde GHSR rs2232165 polimorfizminin MAF değeri 0,02 olarak belirlenmiştir. Bu çalışmada, çağımızda alkol kullanım bozukluğu kadar önemli olan obezite etiyolojisinde de rol oynayan GHSR rs2232165 polimorfizminin Türk erkeklerinde alel frekansı ile ilgili ilk veriler toplanmıştır.Alcohol addiction, which is one of the crucial problems of the current era, arises with the effects of genetic, environmental, culturel, developmental and neurobiological factors. In recent years, the discovery that the neurobiology of eating behavior and the neurobiology of craving for addictive substances have similar characteristics has prompted researchers to investigate the relationship between alcohol addiction and hormones important in food intake such as ghrelin. Even though the importance of the ghrelin system in alcohol use disorder and alcohol craving has been stated by existing studies , more studies are needed to enlighten the biological mechanism of this relationship. Therefore, in this study the relationship between rs2232165 polymorphism in GHSR gene encoding ghrelin receptor and alcohol use disorder in Turkish men with alcohol use problem. Seventy-two individuals diagnosed with alcohol use disorder and 82 healthy men without any substance addiction were included in this study. The GHSR rs2232165 gene polymorphism was genotyped by PCR-RFLP which was optimized for the first time in this study. The allele frequency of the GHSR rs2232165 gene polymorphism was determined in the group with alcohol use problem (n:144) as 0,99 (n:142) for C allele and 0,01 (n:2) for T allele. The frequency of the C allele was 0,98 (n:161) and T allele was 0,02 (n:3) in the healthy group. There was no statistically significant difference between two groups in terms of T allele frequency. Since gene polymorphisms are not affected by environmental factors, individuals in two groups were combined (n=154) in order to determine minor allele frequency (MAF) in Turkish males, and the MAF value of GHSR rs2232165 polymorphism in Turkish males was determined as 0,02. In this study, the first data on the allele frequency of GHSR rs2232165 polymorphism, which plays a role in the etiology of obesity, which is as important as alcohol use disorder in the current era, were collected in Turkish males

The involvement of miRNAs in CYP450 gene expression: A brief review of the literature

Drug biotransformation is a critical process in metabolic elimination of drugs. It occurs mainly in the liver as well as in other tissues such as kidney and small intestine and consists of three stages: Phase I, Phase II and Phase III. Drugs are converted to a more polar and hydrophilic metabolites during Phase I and Phase II, which are excreted by a variety of membrane transporters such as P-glycoprotein and Multidrug Resistance Protein 1 in Phase III. Cytochrome P450 enzyme isoforms achieve 80% of Phase I metabolism. It is well described that there is inter-individual drug response variability such as adverse drug reactions or reduced drug efficacy and that genetic variation within the sequence of biotransformation-related genes affects these different therapeutic outcomes observed between individuals. However, genetic factors cannot completely explain inter-individual differences. Recent studies have shown that epigenetic factors such as histone modification, DNA methylation and non-coding miRNAs heavily influence drug biotransformation through post-transcriptional regulation of metabolism gene expression. It is important to understand the causes of alterations in drug metabolism since varied biotransformation may lead to adverse drug effects or a loss of efficacy. Therefore, in this review, the effects of miRNAs in CYP450 gene expression will be discussed briefly in the light of recent studies

Association of OPRK1 rs963549 and rs997917 polymorphisms with opioid use disorder and related phenotypes

Aim: To evaluate the association between OPRK1 rs963549 and rs997917 and opioid use disorder (OUD) and related phenotypes. Methods: A sample of 208 individuals with (n = 100) and without (n = 108) OUD were enrolled. OPRK1 rs963549 and rs997917 were analyzed by PCR-RFLP. Craving, opioid withdrawal and the intensity of depressive and anxiety symptoms were measured by the appropriate scales. Results:OPRK1 rs963549 variation showed a trend of association with decreased opioid withdrawal. No significant associations were found between OPRK1 rs963549 and rs997917 polymorphisms and craving, depression or anxiety symptoms. Neither single OPRK1 SNPs nor OPRK1 haplotypes were associated with OUD. Conclusion: Our results could be useful for treatment failures of individuals who experience greater opioid withdrawal due to their OPRK1 rs963549 genotypes

Effect of PDYN rs2281285, rs2235749 and rs910080 gene polymorphisms on the intensity of depression symptoms and negative craving in heroin addicts

The present study was undertaken to explore whether prodynorphin (PDYN) polymorphisms have an effect on the intensity of depressive symptoms and negative craving in heroin addicts in a sample of 100 heroin addicts and 108 controls. PDYN rs2281285, rs2225749 and rs910080 polymorphisms were analyzed by PCR-RFLP. Craving and the intensity of depressive symptoms were measured by the Substance Craving Scale and the Beck Depression Inventory-II, respectively. A significant association between depression severity and PDYN rs2281285 (P=0.026) and rs2225749 (P=0.038) polymorphisms was detected. PDYN rs2225749 variation showed a trend association with increased negative craving (P=0.066). We also examined the associations between heroin dependence and PDYN rs2281285, rs2225749 and rs910080 gene polymorphisms at the gene and haplotype levels. The AAA haplotype was more frequent in heroin addicts and shown to be significantly associated with increased risk for heroin dependence (OR, 8.922; 95% CI, 1.116-71.313; P[removed

OPRD1 rs569356 polymorphism has an effect on plasma norbuprenorphine levels and dose/kg-normalized norbuprenorphine values in individuals with opioid use disorder

This study aimed to determine the effects of nine OPRM1, OPRD1 and OPRK1 polymorphisms on plasma BUP and norbuprenorphine (norBUP) concentrations and various treatment responses in a sample of 122 patients receiving BUP/naloxone. Plasma concentrations of BUP and norBUP were detected by LC–MS/MS. PCR-RFLP method was used to genotype polymorphisms. OPRD1 rs569356 GG had significantly lower plasma norBUP concentration (p = 0.018), dose- (p = 0.049) and dose/kg-normalized norBUP values (p = 0.036) compared with AA. Craving and withdrawal symptoms were significantly higher in OPRD1 rs569356 AG+GG relative to AA. There was a statistically significant difference between the OPRD1 rs678849 genotypes in the intensity of anxiety (13.5 for CT+TT and 7.5 for TT). OPRM1 rs648893 TT (18.8 ± 10.8) was significantly different to CC+CT (14.82 ± 11.3; p = 0.049) in view of the intensity of depression. This current study provides the first data on a prominent effect of the OPRD1 rs569356 variation on BUP pharmacology due to its metabolite norBUP. © 2023 Elsevier B.V

Effects of UGT2B7 rs7662029 and rs7439366 polymorphisms on sublingual buprenorphine metabolism in heroin addicts: An improved PCR-RFLP assay for the detection of rs7662029 polymorphism

This study aimed to determine the effects of UGT2B7 rs7662029 and rs7439366 polymorphisms on plasma buprenorphine (BUP) concentration and different treatment responses in a sample of 109 patients with opioid use disorder (OUD) treated with sublingual BUP/naloxone. Polymorphisms were analysed by PCR-RFLP. Plasma concentrations of BUP and its metabolite norbuprenorphine were detected by LC-MS/MS. Craving, withdrawal, depression and anxiety were measured by appropriate scales. OUD patients with rs7439366 CC or rs7662029 GG genotypes had significantly lower dose-normalized (BUP/D) and dose/kg-normalized BUP (BUP/D.kg(-1)) levels than those who were CT or AA carriers. Significant associations between UGT2B7 rs7662029 and increased craving (p = 0.037) and withdrawal symptoms (p = 0.029) were detected. Our findings were pointing to an important role of UGT2B7 in the metabolism of sublingual BUP/naloxone in the heroin addicts for the first time. A novel PCR-RFLP assay was developed for the determination of UGT2B7 rs7662029 polymorphism, based on utilizing novel restriction enzyme